Innate Immune Signaling in Fibrolamellar Carcinoma

纤维板层癌中的先天免疫信号传导

基本信息

批准号：
10707605
负责人：
STEEN HENNING HANSEN
金额：
$ 17.7万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-15 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10707605
关键词：
Ablation Address Affect CASP1 gene CASP8 gene CRISPR/Cas technology Carcinoma Caspase Caspase Inhibitor Catalytic Domain Cell Culture Techniques Cell Death Cells Child Chronic Cyclic AMP-Dependent Protein Kinases DNA Sequence Alteration Data Development Diagnosis Disease Disputes Engineering Exhibits Exons Extrahepatic Family Fibrolamellar Hepatocellular Carcinoma Future Gene Fusion Genes Genomics Heat shock proteins Hepatocarcinogenesis Hepatocyte IL18 gene Immune Immune signaling Immunity Infiltration Inflammasome Inflammation Inflammatory Response Injections Knock-out Link Liver Liver Stem Cell Liver neoplasms Lytic Malignant Neoplasms Malignant neoplasm of liver Mediating Modeling Mus Mutation Natural Immunity Nature Oncogenic Ontology Organoids PRKACA gene Pathogenesis Patients Phenotype Prognosis Proteins Publications Publishing Recurrence Risk Role Signal Pathway Signal Transduction Tail Testing Time Veins Work anakinra curative treatments effective therapy fusion gene genome editing hepatobiliary cancer immune cell infiltrate in vivo inhibitor insight interest liver transplantation mouse model novel preservation prevent programs stem cells tool transcriptome transcriptomic profiling transcriptomics translational study tumor tumorigenesis virtual young adult

项目摘要

Fibrolammellar Carcinoma (FLC) is a rare form of liver cancer that afflicts children and young adults. At the time of diagnosis, the cancer is stage IV in ~60% of patients without effective treatment option. For the remaining 40%, liver transplantation is the only curative therapy, providing a suitable donor can be found in time and even then, with significant risks for short- and long-term complications. Virtually all patients with FLC have a genomic alteration on Chr. 19 resulting in fusion of exon 1 of DNAJB1 of the Hsp40 family to exons 2-10 of PRKACA, a catalytic subunit of PKA. The resulting DNAJB1-PRKACA fusion gene encodes a Dnajb1-Prkaca protein with preserved catalytic activity. Using CRISPR/Cas9 technology combined with hydrodynamic tail vein injection, we and others have shown conclusively that a syntenic Dnajb1- Prkaca fusion gene on mouse Chr. 8 is sufficient to elicit liver cancer with hallmarks of FLC. FLC is thought to arise from liver stem cells, but the mechanisms whereby Dnajb1-Prkaca promotes oncogenic transformation are not well understood. To permit functional studies in a developmentally relevant context, we used CRISPR/Cas9 technology to engineer syntenic Dnajb1-Prkaca in mouse liver organoids. We discovered that liver organoids expressing Dnajb1-Prkaca are prone to undergoing lytic cell death. Next, we performed extensive transcriptome analyses that revealed an innate immune signature prominently featuring constituents of the NLRC4 inflammasome. We moreover determined that Dnajb1-Prkaca expressing liver organoids exhibit significantly elevated levels of cleaved caspase-8 consistent with its potent activation. In this work, we will first test if the NLRC4 inflammasome is activated by Dnajb1-Prkaca in mouse liver organoids and establish its role in lytic cell death. We will then determine if the Nlrc4 and/or Casp-8 genes are necessary for tumorigenesis elicited by engineering of the Dnajb1-Prkaca fusion gene in the mouse liver. The latter would represent a major breakthrough in understanding the pathogenesis of FLC.

纤维板层癌 (FLC) 是一种罕见的肝癌，困扰儿童和年轻人。当时诊断后，约 60% 的患者癌症处于 IV 期，而没有有效的治疗选择。对于剩余的 40%，肝移植是唯一的治疗方法，只要能及时找到合适的供体，甚至然后，存在短期和长期并发症的巨大风险。几乎所有 FLC 患者的 Chr. 基因组均发生改变。 19 导致 DNAJB1 的外显子 1 融合 Hsp40 家族与 PRKACA（PKA 的催化亚基）的外显子 2-10 相连接。由此产生的 DNAJB1-PRKACA 融合基因编码具有保留催化活性的 Dnajb1-Prkaca 蛋白。使用CRISPR/Cas9技术结合流体动力尾静脉注射，我们和其他人已经最终证明同线 Dnajb1- 小鼠 Chr. Prkaca 融合基因8 足以诱发具有 FLC 特征的肝癌。 FLC被认为起源于肝干细胞，但Dnajb1-Prkaca促进致癌的机制转型还没有被很好的理解。为了允许在与发育相关的背景下进行功能研究，我们使用 CRISPR/Cas9 技术在小鼠肝脏类器官中设计同线 Dnajb1-Prkaca。我们发现表达 Dnajb1-Prkaca 的肝脏类器官容易发生裂解性细胞死亡。接下来我们表演了广泛的转录组分析揭示了先天免疫特征的显着特征 NLRC4 炎症小体。我们还确定表达 Dnajb1-Prkaca 的肝脏类器官表现出裂解的 caspase-8 水平显着升高，与其有效激活一致。在这项工作中，我们将首先测试小鼠肝脏类器官中的 NLRC4 炎症小体是否被 Dnajb1-Prkaca 激活并确定其在裂解细胞死亡中的作用。然后我们将确定 Nlrc4 和/或 Casp-8 基因是否是必需的用于小鼠肝脏中 Dnajb1-Prkaca 融合基因工程引发的肿瘤发生。后者将代表了理解 FLC 发病机制的重大突破。